CN111621527A - Biological fermentation process and device thereof - Google Patents

Biological fermentation process and device thereof Download PDF

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Publication number
CN111621527A
CN111621527A CN202010400222.8A CN202010400222A CN111621527A CN 111621527 A CN111621527 A CN 111621527A CN 202010400222 A CN202010400222 A CN 202010400222A CN 111621527 A CN111621527 A CN 111621527A
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fermentation
tank
culture
culture medium
seed
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臧武波
程秋洁
彭荣达
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Zhejiang Lanbo Biotechnology Co ltd
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Zhejiang Lanbo Biotechnology Co ltd
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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P1/00Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12MAPPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M27/00Means for mixing, agitating or circulating fluids in the vessel
    • C12M27/02Stirrer or mobile mixing elements
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12MAPPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M29/00Means for introduction, extraction or recirculation of materials, e.g. pumps
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12MAPPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/02Means for regulation, monitoring, measurement or control, e.g. flow regulation of foam
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12MAPPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/12Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
    • C12M41/18Heat exchange systems, e.g. heat jackets or outer envelopes
    • C12M41/22Heat exchange systems, e.g. heat jackets or outer envelopes in contact with the bioreactor walls

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  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Wood Science & Technology (AREA)
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  • Bioinformatics & Cheminformatics (AREA)
  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
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  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

The invention provides a biological fermentation process and a device thereof, belonging to the technical field of biology. The method solves the technical problems of complex production process, low production efficiency, high production cost and the like of biological fermentation in the prior art. The biological fermentation process and the device thereof are characterized in that the fermentation process comprises the following steps: the method comprises the following steps: preparation work: (1) identifying the purity and the activity of the strain; (2) preparing a fermentation culture medium, wherein the fermentation culture medium comprises the following components: 35g/L of carbon source, 6g/L of yeast powder, 2g/L of diammonium hydrogen phosphate, 1g/L of monopotassium phosphate, 0.4g/L of magnesium sulfate and 0.2g/L of defoaming agent; step two: activating strains and culturing seeds; step three: fermenting in a seed tank; step four: and (3) a main fermentation stage. The invention has the advantages of simple process and high production efficiency.

Description

Biological fermentation process and device thereof
Technical Field
The invention belongs to the technical field of biology, and relates to a fermentation process, in particular to a biological fermentation process.
Background
The biological fermentation engineering is an important component of biological engineering, microorganisms utilize carbohydrate to ferment and produce various industrial solvents and chemical raw materials, and can be cultured in a large scale so as to fully exert the inherent potential of the microorganisms and provide great economic benefit and social benefit for people.
Disclosure of Invention
The invention aims to solve the problems in the prior art and provides a biological fermentation process and a device thereof.
The purpose of the invention can be realized by the following technical scheme: a biological fermentation process and a device thereof are characterized in that the fermentation process comprises the following steps:
the method comprises the following steps: preparation work:
(1) identifying the purity and the activity of the strain;
(2) preparing a fermentation culture medium, wherein the fermentation culture medium comprises the following components: 35g/L of carbon source, 6g/L of yeast powder, 2g/L of diammonium hydrogen phosphate, 1g/L of monopotassium phosphate, 0.4g/L of magnesium sulfate and 0.2g/L of defoaming agent;
step two: activating strains and culturing seeds:
(1) activating the frozen and preserved strain in a slant, and purifying the strain in a flat plate to finally obtain a strain slant;
(2) a shake flask culture stage, in which a ring of purified strains are taken for shake flask culture;
step three: fermenting in a seeding tank:
(1) checking the seed tank to ensure the inside of the tank to be clean and prevent impurities from being generated after feeding; a source of water of in-tank volume 1/3 is required in the tank to avoid damage to the bearing connections;
(2) batching in a seeding tank, detecting the pH value of the seeding tank, and controlling the pH value of the fermentation in the seeding tank to be 6.5-7.5;
(3) sequentially carrying out sterilization, cooling, inoculation and fermentation of a seed tank;
step four: and (3) a main fermentation stage:
(1) batching in a fermentation tank, detecting the pH value of the fermentation tank, and controlling the pH value of the fermentation tank to be 6.0-7.0;
(2) sterilizing a fermentation tank, namely heating a jacket to 90 ℃, directly heating steam to 121 ℃, keeping the temperature of 0.12MPa for 20 minutes, and then cooling and culturing the fermentation tank, wherein the fermentation tank culture stage is as follows: the temperature of the fermentation tank is 26-30 ℃, the pressure of the fermentation tank is 0.05MPa, the air volume is 0.2-0.6VVM, the stirring speed is 150-250rpm, the dissolved oxygen is more than 30 percent, the culture time is 15-20 hours, and the culture is stored at normal temperature and normal pressure after the completion.
The shake flask culture steps are as follows:
(1) inoculating into a 250mL triangular flask containing 20mL seed culture medium, and culturing in a shaking table;
(2) respectively taking 1mL of seed solution, inoculating the seed solution into five 250mL triangular flasks containing 20mL of fermentation medium, and culturing in a shaking table; the inoculation amount is 45 percent of the actual culture medium loading amount of the triangular flask, and the PH value is controlled to be 7.2-7.6.
The seed tank in the third step comprises a tank body, wherein the upper end and the lower end of the tank body are both hemispherical, the upper part of the tank body is provided with a feeding structure, the lower part of the tank body is provided with a discharging structure, the tank body is provided with a stirring mechanism and a defoaming structure, the stirring mechanism comprises a stirring shaft, an auxiliary frame and a plurality of stirring sheets, the auxiliary frame is fixed in the tank body, one end of the stirring shaft is rotatably arranged on the tank body through a bearing, the other end of the stirring shaft is rotatably arranged on the auxiliary frame through a bearing, and the stirring shaft is also connected with a power structure; the defoaming structure further comprises a defoaming disc and an auxiliary plate, the defoaming disc is fixed on the stirring shaft, a plurality of defoaming blocks are fixed on the defoaming disc, the auxiliary plate is fixed on the tank body, and the auxiliary plate is further fixed with auxiliary blocks.
Structure more than adopting, power structure drive the (mixing) shaft and rotate, and the stirring piece stirs jar internal raw materials, through setting up the auxiliary frame for the (mixing) shaft rotates stably, sets up the defoaming dish simultaneously on the (mixing) shaft, under the cooperation of accessory plate, defoaming piece and accessory block, carries out the processing to the foam, prevents that the foam from spilling over, has excellent in use effect's advantage.
The external side of jar be provided with and be used for refrigerated cooling chamber, the intercommunication has cooling water inlet and cooling water outlet on the cooling chamber, cooling water inlet is connected with a inlet tube, the cooling water outlet is connected with a water tank.
Structure more than adopting, the cooling water gets into the cooling chamber through the cooling water import, cools down the jar body, improves the efficiency of fermentation.
The power structure include mounting bracket, motor power and gearbox, the mounting bracket is fixed on the jar body, motor power and gearbox are all fixed on the mounting bracket, motor power's output shaft sets up vertically downwards, the output of gearbox is connected with motor power's output shaft, the output of gearbox is connected with the (mixing) shaft.
The feed structure include liquid inlet pipe, liquid feeder hopper and fixed inlet pipe, the liquid inlet pipe is fixed on the jar body, the one end of liquid inlet pipe stretches into jar body lower part, the other end and the liquid feeder hopper of liquid inlet pipe are linked together, the solid inlet pipe is fixed on the jar body, and solid inlet pipe and jar body upper portion part communicate.
Structure more than adopting, the one end of liquid inlet pipe stretches into jar body lower part, and liquid raw materials can directly get into jar body lower part for mix more high-efficient.
The discharging structure comprises a discharging pipe and a valve, the discharging pipe is arranged on the tank body and communicated with the tank body, and the valve is arranged on the discharging pipe.
And an auxiliary sheet for accelerating discharging is fixed on the stirring shaft.
By adopting the structure and arranging the auxiliary pieces, when discharging, the auxiliary blocks rotate to accelerate the discharging speed.
The tank body is also provided with a vent pipe, the vent pipe is connected with an air pump, and the vent pipe is provided with a plurality of air outlets.
By adopting the structure, the strains can contact more air by arranging the air outlets.
The tank body is also provided with a temperature detector for measuring the temperature in the tank body.
Compared with the prior art, the invention has the following advantages: simple process and high production efficiency.
Drawings
Fig. 1 is a schematic cross-sectional structure of the present invention.
In the figure, 1, a tank body; 2. a stirring shaft; 3. an auxiliary frame; 4. a stirring sheet; 5. a bearing; 6. a defoaming plate; 7. an auxiliary plate; 8. defoaming blocks; 9. an auxiliary block; 10. a cooling chamber; 11. a cooling water inlet; 12. a cooling water outlet; 13. a mounting frame; 14. a power motor; 15. a gearbox; 16. a liquid feed conduit; 17. a liquid feed hopper; 18. fixing a feeding pipe; 19. a discharge pipe; 20. a valve; 21. an auxiliary sheet; 22. a breather pipe; 22a and an air outlet hole; 23. an air pump; 24. a temperature detector.
Detailed Description
The following are specific embodiments of the present invention and are further described with reference to the drawings, but the present invention is not limited to these embodiments.
As shown in FIG. 1, the present biological fermentation process and apparatus thereof, in this embodiment, the fermentation process comprises the following steps:
the method comprises the following steps: preparation work:
(1) identifying the purity and the activity of the strain;
(2) preparing a fermentation culture medium, wherein the fermentation culture medium comprises the following components: 35g/L of carbon source, 6g/L of yeast powder, 2g/L of diammonium hydrogen phosphate, 1g/L of monopotassium phosphate, 0.4g/L of magnesium sulfate and 0.2g/L of defoaming agent;
step two: activating strains and culturing seeds:
(1) activating the frozen and preserved strain in a slant, and purifying the strain in a flat plate to finally obtain a strain slant;
(2) a shake flask culture stage, in which a ring of purified strains are taken for shake flask culture;
step three: fermenting in a seeding tank:
(1) checking the seed tank to ensure the inside of the tank to be clean and prevent impurities from being generated after feeding; a source of water of in-tank volume 1/3 is required in the tank to avoid damage to the bearing connections;
(2) batching in a seeding tank, detecting the pH value of the seeding tank, and controlling the pH value of the fermentation in the seeding tank to be 6.5-7.5;
(3) sequentially carrying out sterilization, cooling, inoculation and fermentation of a seed tank;
step four: and (3) a main fermentation stage:
(1) batching in a fermentation tank, detecting the pH value of the fermentation tank, and controlling the pH value of the fermentation tank to be 6.0-7.0;
(2) sterilizing a fermentation tank, namely heating a jacket to 90 ℃, directly heating steam to 121 ℃, keeping the temperature of 0.12MPa for 20 minutes, and then cooling and culturing the fermentation tank, wherein the fermentation tank culture stage is as follows: the temperature of the fermentation tank is 26-30 ℃, the pressure of the fermentation tank is 0.05MPa, the air volume is 0.2-0.6VVM, the stirring speed is 150-250rpm, the dissolved oxygen is more than 30 percent, the culture time is 15-20 hours, and the culture is stored at normal temperature and normal pressure after the completion.
The shake flask culture steps are as follows:
(1) inoculating into a 250mL triangular flask containing 20mL seed culture medium, and culturing in a shaking table;
(2) respectively taking 1mL of seed solution, inoculating the seed solution into five 250mL triangular flasks containing 20mL of fermentation medium, and culturing in a shaking table; the inoculation amount is 45 percent of the actual culture medium loading amount of the triangular flask, and the PH value is controlled to be 7.2-7.6.
The seed tank in the third step comprises a tank body 1, wherein the upper end and the lower end of the tank body 1 are both hemispherical, the upper part of the tank body 1 is provided with a feeding structure, the lower part of the tank body 1 is provided with a discharging structure, the tank body 1 is provided with a stirring mechanism and a defoaming structure, the stirring mechanism comprises a stirring shaft 2, an auxiliary frame 3 and a plurality of stirring sheets 4, in the embodiment, the number of the stirring sheets 4 is twelve, the auxiliary frame 3 is fixed in the tank body 1, one end of the stirring shaft 2 is rotatably arranged on the tank body 1 through a bearing 5, the other end of the stirring shaft 2 is rotatably arranged on the auxiliary frame 3 through the bearing 5, and the stirring shaft 2 is also connected with a power structure; the defoaming structure also comprises a defoaming disc 6 and an auxiliary plate 7, wherein the defoaming disc 6 is fixed on the stirring shaft 2, a plurality of defoaming blocks 8 are fixed on the defoaming disc 6, in the embodiment, the number of the defoaming blocks 8 is four, the auxiliary plate 7 is fixed on the tank body 1, and the auxiliary plate 7 is also fixed with an auxiliary block 9.
Structure more than adopting, power structure drive (mixing) shaft 2 rotates, and stirring piece 4 stirs the raw materials of jar internal 1, through setting up auxiliary frame 3 for (mixing) shaft 2 rotates stably, sets up defoaming dish 6 simultaneously on (mixing) shaft 2, under the cooperation of accessory plate 7, defoaming piece 8 and auxiliary block 9, carries out the processing to the foam, prevents that the foam from spilling over, has excellent in use effect's advantage.
The outer side of the tank body 1 is provided with a cooling chamber 10 for cooling, the cooling chamber 10 is communicated with a cooling water inlet 11 and a cooling water outlet 12, the cooling water inlet 11 is connected with a water inlet pipe, and the cooling water outlet 12 is connected with a water tank.
Structure more than adopting, the cooling water passes through cooling water inlet 11 and gets into cooling chamber 10, cools down jar body 1, improves the efficiency of fermentation.
The power structure includes mounting bracket 13, motor power 14 and gearbox 15, and mounting bracket 13 is fixed on jar body 1, and motor power 14 and gearbox 15 are all fixed on mounting bracket 13, and motor power 14's output shaft is vertical sets up downwards, and the output of gearbox 15 is connected with motor power 14's output shaft, and the output of gearbox 15 is connected with (mixing) shaft 2.
The feeding structure comprises a liquid feeding pipe 16, a liquid feeding hopper 17 and a fixed feeding pipe 18, wherein the liquid feeding pipe 16 is fixed on the tank body 1, one end of the liquid feeding pipe 16 extends into the lower part of the tank body 1, the other end of the liquid feeding pipe 16 is communicated with the liquid feeding hopper 17, the solid feeding pipe is fixed on the tank body 1, and the solid feeding pipe is communicated with the upper part of the tank body 1.
Structure more than adopting, the one end of liquid inlet pipe 16 stretches into jar body 1 part down, and liquid raw materials can directly get into jar body 1 part down for mix more high-efficient.
The discharging structure comprises a discharging pipe 19 and a valve 20, the discharging pipe 19 is arranged on the tank body 1, the discharging pipe 19 is communicated with the tank body 1, and the valve 20 is arranged on the discharging pipe 19.
An auxiliary sheet 21 for accelerating discharging is fixed on the stirring shaft 2.
By adopting the structure, the auxiliary piece 21 is arranged, and the auxiliary block 9 rotates to accelerate the discharging speed during discharging.
The tank body 1 is further provided with a vent pipe 22, the vent pipe 22 is connected with an air pump 23, and the vent pipe 22 is provided with a plurality of air outlets 22a, in this embodiment, the number of the air outlets 22a is eleven.
With the above structure, the seed can contact more air by opening the air outlet hole 22 a.
The tank body 1 is also provided with a temperature detector 24 for measuring the temperature in the tank body 1.
The specific embodiments described herein are merely illustrative of the spirit of the invention. Various modifications or additions may be made to the described embodiments or alternatives may be employed by those skilled in the art without departing from the spirit or ambit of the invention as defined in the appended claims.

Claims (2)

1. A biological fermentation process and a device thereof are characterized in that the fermentation process comprises the following steps:
the method comprises the following steps: preparation work:
(1) identifying the purity and the activity of the strain;
(2) preparing a fermentation culture medium, wherein the fermentation culture medium comprises the following components: 35g/L of carbon source, 6g/L of yeast powder, 2g/L of diammonium hydrogen phosphate, 1g/L of monopotassium phosphate, 0.4g/L of magnesium sulfate and 0.2g/L of defoaming agent;
step two: activating strains and culturing seeds:
(1) activating the frozen and preserved strain in a slant, and purifying the strain in a flat plate to finally obtain a strain slant;
(2) a shake flask culture stage, in which a ring of purified strains are taken for shake flask culture;
step three: fermenting in a seeding tank:
(1) checking the seed tank to ensure the inside of the tank to be clean and prevent impurities from being generated after feeding; a source of water of in-tank volume 1/3 is required in the tank to avoid damage to the bearing connections;
(2) batching in a seeding tank, detecting the pH value of the seeding tank, and controlling the pH value of the fermentation in the seeding tank to be 6.5-7.5;
(3) sequentially carrying out sterilization, cooling, inoculation and fermentation of a seed tank;
step four: and (3) a main fermentation stage:
(1) batching in a fermentation tank, detecting the pH value of the fermentation tank, and controlling the pH value of the fermentation tank to be 6.0-7.0;
(2) sterilizing a fermentation tank, namely heating a jacket to 90 ℃, directly heating steam to 121 ℃, keeping the temperature of 0.12MPa for 20 minutes, and then cooling and culturing the fermentation tank, wherein the fermentation tank culture stage is as follows: the temperature of the fermentation tank is 26-30 ℃, the pressure of the fermentation tank is 0.05MPa, the air volume is 0.2-0.6VVM, the stirring speed is 150-250rpm, the dissolved oxygen is more than 30 percent, the culture time is 15-20 hours, and the culture is stored at normal temperature and normal pressure after the completion.
2. The biological fermentation process and apparatus thereof according to claim 1, wherein the shake flask cultivation step is:
(1) inoculating into a 250mL triangular flask containing 20mL seed culture medium, and culturing in a shaking table;
(2) respectively taking 1mL of seed solution, inoculating the seed solution into five 250mL triangular flasks containing 20mL of fermentation medium, and culturing in a shaking table; the inoculation amount is 45 percent of the actual culture medium loading amount of the triangular flask, and the PH value is controlled to be 7.2-7.6.
CN202010400222.8A 2020-05-12 2020-05-12 Biological fermentation process and device thereof Pending CN111621527A (en)

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CN202010400222.8A CN111621527A (en) 2020-05-12 2020-05-12 Biological fermentation process and device thereof

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Application Number Priority Date Filing Date Title
CN202010400222.8A CN111621527A (en) 2020-05-12 2020-05-12 Biological fermentation process and device thereof

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Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN203517948U (en) * 2013-10-25 2014-04-02 罗依依 Smoke exhaust ventilator with smoke gas purification and grease recovery function
CN106085828A (en) * 2016-07-31 2016-11-09 盐城市大明化工机械有限公司 Seed tank with agitating device
CN108239602A (en) * 2017-12-04 2018-07-03 连云港巨森农业科技发展有限公司 A kind of production technology of complex microbial inoculum
CN208878475U (en) * 2018-07-25 2019-05-21 成都道盛路通新材料有限公司 A kind of novel normal-temperature asphalt modifier blend tank
CN110559917A (en) * 2019-09-26 2019-12-13 虞伟明 a mixing apparatus that is used for discharging of dairy products production and processing efficient
CN110982860A (en) * 2019-12-10 2020-04-10 山东天智绿业生物科技有限公司 Fermentation method of curdlan

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN203517948U (en) * 2013-10-25 2014-04-02 罗依依 Smoke exhaust ventilator with smoke gas purification and grease recovery function
CN106085828A (en) * 2016-07-31 2016-11-09 盐城市大明化工机械有限公司 Seed tank with agitating device
CN108239602A (en) * 2017-12-04 2018-07-03 连云港巨森农业科技发展有限公司 A kind of production technology of complex microbial inoculum
CN208878475U (en) * 2018-07-25 2019-05-21 成都道盛路通新材料有限公司 A kind of novel normal-temperature asphalt modifier blend tank
CN110559917A (en) * 2019-09-26 2019-12-13 虞伟明 a mixing apparatus that is used for discharging of dairy products production and processing efficient
CN110982860A (en) * 2019-12-10 2020-04-10 山东天智绿业生物科技有限公司 Fermentation method of curdlan

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Application publication date: 20200904